Cell lysates were collected in RIPA lysis buffer (1% Triton-X-100, 20 mM Tris, pH 7.5, 137 mM NaCl, 1 mM EGTA, 10% glycerol, 1.5 mM MgCl2, and protease inhibitor mixture and phosphatase inhibitors; latter 2 were from Roche). Lysates were sonicated and centrifuged at 4 °C. Per lane, whole-cell lysate was separated on 12% SDS-acrylamide gels and transferred on Immobilon PVDF membranes (Millipore). The membranes were probed with primary antibodies overnight at 4 °C and incubated for 1 h with secondary peroxidase-conjugated antibodies (CST). Chemiluminescent signals were then developed with Lumiglo reagent (Cell Signaling Technology) and detected by the ChemiDoc XRS gel documentation system (Bio-rad). Antibodies include anti-Ube2v1 (Abcam, monoclonal ab151725), anti-E-cadherin (Santa Cruz, monoclonal sc-21791), anti-N-cadherin (Boster, polyclonal BA0673), anti-Fibronectin (Boster, polyclonal BA1771), anti-Vimentin (Abcam, monoclonal, ab8978), anti-β-Catenin (Cell Signaling Technology, monoclonal #8480), anti-Twist1 (Proteintech, 25465), anti-Snai1 (Bioss, bs-2441R), anti-LC3B (CST, monoclonal 3868), anti-SQSTM1/p62 (CST, polyclonal 5114), anti-H4K16ac (Immunoway, polyclonal YM3317), anti-Beclin1 (Boster, polyclonal PB0014), anti-histone H3 (Abcam, polyclonal ab1791), and anti-Sirt1 (Santa Cruz, monoclonal sc-74504).
Anti beclin1
Manufactured by Boster Bio
Sourced in China
Anti-Beclin1 is a laboratory reagent that can be used to detect the Beclin-1 protein, which is involved in the regulation of autophagy, a cellular process that recycles damaged or unwanted components. The reagent can be used in various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of Beclin-1 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti histone h3, by Abcam (1 mentions)
Anti sirt1, by Santa Cruz Biotechnology (1 mentions)
Anti vimentin, by Abcam (1 mentions)
Chemidoc xrs gel documentation system, by Bio-Rad (1 mentions)
Lumiglo reagent, by Cell Signaling Technology (1 mentions)
Anti β catenin, by Cell Signaling Technology (1 mentions)
Immobilon pvdf membrane, by Merck Group (1 mentions)
Anti twist1, by Proteintech (1 mentions)
Anti fibronectin, by Boster Bio (1 mentions)
Anti e cadherin, by Santa Cruz Biotechnology (1 mentions)
Pd37 type microscope, by Olympus (1 mentions)
Anti ampk, by Abcam (1 mentions)
Dab horseradish peroxidase color development kit, by Boster Bio (1 mentions)
Anti p62, by Boster Bio (1 mentions)
Anti mtor, by Abcam (1 mentions)
2 protocols using anti beclin1
1
Western Blot Analysis for EMT and Autophagy
2
Immunohistochemical Analysis of Autophagy Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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After the paraffin-embedded sections were dewaxed, we transferred the slides to 100% alcohol, 95% alcohol, 75% alcohol, and ddH 2 O 2 min twice respectively. After antigen retrieval, put them in hydrogen peroxide (3%) for 10 min to eliminate endogenous peroxidase activity. After washing with PBS three times, the sections of different groups were incubated with anti-AMPK (1:100, abcam, Cambridge, UK), anti-mTOR (1:400, abcam, Cambridge, UK), anti-ULK1 (1:300, Boster, Wuhan, Hubei, China), anti-Beclin-1 (1:200, Boster, Wuhan, Hubei, China), anti-p62 (1:200, Boster, Wuhan, Hubei, China), anti-LC3-II/LC3-I (1:100, Boster, Wuhan, Hubei, China) at 37 for 1 h. After washing with PBS three times, the secondary antibody (Boster) was added and immunostaining was performed using a DAB horseradish peroxidase color development kit (Boster, Wuhan, Hubei, China), and then, sections were counterstained with hematoxylin and made transparent with xylene.
Finally, sections were observed with the PD37 type microscope (Olympus, Japan). Under 20× magnification, pictures were taken in five random fields. The catalog numbers: anti-ULK1 (A00584-1), anti-Beclin-1 (PB9076), anti-LC3-II/LC3-I (BM4827), anti-p62 (PB0458), anti-AMPK (ab32047), and anti-mTOR (ab32028).
Finally, sections were observed with the PD37 type microscope (Olympus, Japan). Under 20× magnification, pictures were taken in five random fields. The catalog numbers: anti-ULK1 (A00584-1), anti-Beclin-1 (PB9076), anti-LC3-II/LC3-I (BM4827), anti-p62 (PB0458), anti-AMPK (ab32047), and anti-mTOR (ab32028).
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